The ChromoTek Spot-Trap® is an affinity matrix for immunoprecipitation of Spot-tagged proteins.
It consists of an anti-Spot-Tag Nanobody/ VHH, which is conjugated to beads.
More information about the Spot capture and detection tag can be found here.
Immunoprecipitation (IP)/ Co-IP
IP/ Co-IP for mass spec (MS) analysis
For protein purification use Spot-Cap
When use Spot-Trap? When use Spot-Cap? Click here.
Spot-Tag is an inert 12 amino acid peptide tag with the sequence PDRVRAVSHWSS.
anti-Spot VHH, Spot binding protein, Spot Nanobody, or anti-Spot single domain antibody fragment (sdAb)
Benefit from Spot-Trap for Immunoprecipitation (IP)
- Reduced background
- High binding affinity, low dissociation constant KD of 6 nM for Spot fusion proteins
- Harsh washing conditions
- High reproducibility
- Short incubation time of 30-60 minutes
- Extraordinary low background compared to other affinity resins
Comparison of matrices & properties
We offer Spot-Trap as Agarose, Magnetic Agarose, and Magnetic Particles M-270.
|Spot-Trap Agarose||Spot-Trap Magnetic Agarose||Spot-Trap Magnetic Particles M-270|
|Size Spot-tagged protein*||Small to large||Small to large||Small to very large|
* Does depend on protein size and shape, protein multimers, complexes and interaction partners
What Spot-Trap shall I use?
- Spot-Trap Agarose, when lowest background and high binding capacity IP is needed.
- Spot-Trap Magnetic Agarose, when magnetic separation and high binding capacity IP is needed.
- Spot-Trap Magnetic Beads, when very large proteins/complexes are investigated, and magnetic separation is needed for IP.
- iST Spot-Trap kit for immunoprecipitation plus sample preparation for mass spectrometry (MS)
Spot-Trap binds to the sequence motif PDRVRAVSHWSS.
Spot-Trap is a very low background affinity reagent. See benchmark of Spot-Trap Agarose and Magnetic Agarose with other suppliers’ capture tools. Immunoprecipitations were conducted from HEK293T cell lysates without Spot-Tag present and normalized for equal binding capacity. The IPs were done according to respective manufacturers’ protocols. It is apparent that Spot-Trap does not only lack heavy and light chain antibody contaminations but also unspecific binding. This makes Spot-Trap a superior affinity capture tool for single band purification of Spot-tagged proteins.
ChromoTek’s proprietary and novel Spot-System is the first peptide-tag specific Nanobody for universal capture & detection applications. It comprises Spot-Tag®, an inert 12 amino acid peptide-tag (PDRVRAVSHWSS), which has been affinity optimized, and the universal, rugged Spot-Nanobody that specifically binds to Spot-tagged proteins with high affinity.
Spot-Tag peptide is derived from the protein beta-catenin: its sequence has been optimized for higher affinity binding to the Spot-Tag Nanobody. For further information see here.
Application notes & posters
Spot-Label samples available
Only for research applications, not for diagnostic or therapeutic use!